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Allozyme variation in some marine mussel species at anthropogenic stress and normal conditions Nikiforov S.M. THE PURPOSE: to compare some parameters of allozyme variation in natural settlements of marine mollusks from unpolluted areas with the same in areas where settlements are under stress by anthropogenic pollution. Table 1. Content of trace metals in water ((mkg/l) above line) and bottom sediments ((mkg/g) below line) in some part of Peter the Great Bay. Area Amurski Bay Nakhodka Bay Zn 2.9-3.9 Cu Cd Ni 0.58-2.90 0.08-0.20 0.60-0.80 81550** - 5-109** - 0.205.48** - 1-136** - 54-163* 16-66* 0.9-1.3 18-42** 0.080.20-0.64 0.02-0.11 0.30-0.38 FESMR 0.48 0-96 0-16 0.0-4.5 0-34* * – content are higher then “threshold effects levels” ** – content are higher then “probable effects levels” Table 2. Content of organic pollutants in water ((mkg/l) above line) and bottom sediments ((mkg/g) below line) in some part of Peter the Great Bay. Район Амурский залив зал. Находка НУВ 40-60 0.032.72** 0-588 ФЕН 4 - СПАВ 50 - 10 18705200 - ДДТ 3.2 0.526.0** - ГХЦГ 1.8 0.2-1.3 - 0.14*0.81* 0-152 0-4 0.006ДВГМЗ 0.011 1.4-15.6* 4.44-5.70 0.000.25* * – content are higher then “threshold effects levels” ** – content are higher then “probable effects levels” НУВ – petroleum hydrocarbons, ФЕН – phenols, СПАВ – anionic detergents, ДДТ – total contents DDT and it metabolites (DDD and DDE (water - ng/l, sediments – ng/g dry weight). ГХЦГ – – total contents organochlorines (water - ng/l, sediments – ng/g dry weigh). Fig. 1. Sites of mussels collection in Peter the Great Bay Fig. 2. Sites of mussels collection in polluted (black circles) and unpolluted (light circles) areas in Peter the Great Bay Table 3.The statistical significance of differences in allele frequencies (Р) among Mytilus trossulus settlements from polluted and unpolluted sites in Nakhodka Bay, Vostok Bay, Amurski Bay and Gould Horn Bay. Locus Ааt-1 Ааt-2 Gpi Idh-2 Lap Mdh-1 Ме-1 Ме-2 Оcdh Iph Pgm 6-Pdh Est-1 Est-2 Est-3 Nakhodka н.с. н.с. <0.05 н.с. н.с. н.с. н.с. н.с. н.с. н.с. н.с. н.с. <0.05 н.с. н.с. Vostok н.с. <0.01 н.с. н.с. н.с. н.с. н.с. <0.05 <0.001 <0.001 н.с. н.с. н.с. Gould horn н.с. н.с. н.с. <0.01 н.с. <0.01 н.с. 0.05 н.с. <0.001 н.с. <0.001 н.с. <0.01 н.с. н.с. – the differences are not significant (P> 0.05). Amurski н.с. н.с. <0.01 н.с. н.с. н.с. н.с. н.с. н.с. <0.001 н.с. <0.001 н.с. н.с. н.с. Table 4. Nei’s indexes of genetic similarity among Mytilus trossulus settlements from polluted and unpolluted sites in Nakhodka Bay, Vostok Bay,Amurski Bay and Gould Horn Bay. Locus Ааt-1 Ааt-2 Gpi Idh-1 Lap Мdh-1 Ме-1 Ме-2 Оcdh Iph Pgm 6-Pdh Est-1 Est-2 Est-3 Mean Nakhodka Bay 0.998 0.999 0.934 0.994 0.956 0.999 0.999 0.999 0.989 0.992 0.998 0.999 0.860 0.996 0.997 0.983 Vostok Bay 0.984 0.976 0.969 1.000 0.999 0.998 0.994 0.923 0.869 0.799 0.990 0.981 0.970 0.958 Gould Horn Bay 0.999 1.000 0.992 0.986 0.992 0.995 0.999 0.986 0.997 0.962 0.992 0.957 0.982 0.944 0.979 0.985 Amurski Bay 0.999 1.000 0.882 0.992 0.974 1.000 0.997 0.999 0.999 0.920 0.969 0.919 0.977 0.991 0.967 0.930 Table 5. The statistical significance of differences in observed heterozygosity (P) among Mytilus trossulus settlements from polluted and unpolluted sites in Nakhodka Bay, Vostok Bay,Amurski Bay and Gould Horn Bay. Locus Nakhodka Vostok Bay Gould Horn Bay Bay н.с. н.с. Ааt-1 н.с. н.с. Ааt-2 <0.05 н.с. н.с. Gpi н.с. <0.05 н.с. Idh-1 н.с. <0.01 н.с. Lap н.с. н.с. н.с. Мdh-1 н.с. н.с. н.с. Ме-1 н.с. н.с. н.с. Ме-2 н.с. н.с. н.с. Оcdh н.с. н.с. <0.01 Iph н.с. <0.05 н.с. Pgm н.с. н.с. н.с. 6-Pdh н.с. н.с. н.с. Est-1 н.с. <0.01 н.с. Est-2 н.с. <0.01 н.с. Est-3 н.с. н.с. н.с. Mean н.с. – the differences are not significant (P> 0.05). Amurski Bay н.с. н.с. н.с. <0.05 н.с. н.с. н.с. н.с. н.с. н.с. <0.05 н.с. н.с. н.с. н.с. н.с. Table 6. 5-aminolivulinate (ALA), aminolivulin syntase (ALA-syntase), protoporphyrin (PP), aminopyrena-Ndemethylase (APND), cytochromes Cyt.(a+a1), Cyt.b, Cyt.(c+c1), Cyt.P450 activity at mussels tissue from polluted (Gaidamak Bay) and unpolluted (Srednyaya Bay) sites in Vostok Bay. Site Srednyaya Bay Gaidamak Bay n 8 ALA* 1196212 ALA-Syntase 28650 PP 5613 APND* 19324 8 1463270 35773 9427 37932 Cyt. (a+a3) 8 13116 Cyt. b 1219 Cyt. (c+c1)* 16817 Cyt. P450* 20625 Srednyaya Bay 8 Gaidamak 15824 16816 23628 39646 Bay ALA – нмоль/мг белка; ALA-syntase – синтез пмоль ALA мг/белка/мин; РР – нг/мг белка; APND – пмоль мг/белка/ мин; Cyt. (a+a3), Cyt. b, Cyt. (c+c1), Cyt. P450 – пмоль/мг белка. n – число исследованных особей; * the differences are statistically significant (Р<0.05). Table 7. The statistical significance of differences in allele frequencies (Pa) and observed heterozygosity (Ph) among Tegula rustica settlements from polluted and unpolluted sites in Peter the Great Bay. Locus Ааt Df Fdh Gpi Gpt Idh Мue Pgm Ра <0.05 <0.05 н.с. <0.001 <0.05 н.с. н.с. н.с. Рh н.с. <0.05 н.с. н.с. н.с. н.с. <0.001 н.с. Table 8. The statistical significance of differences in allele frequencies (Pa) and observed heterozygosity (Ph) among Ischnochiton hakodadensis settlements from polluted and unpolluted sites in Peter the Great Bay. Locus Ааt Df Idh-2 Acph Lap-1 Lap-2 Ме-1 Iph Est-2 Mue Ра <0.001 н.с. н.с. <0.001 <0.001 н.с. <0.001 <0.005 <0.01 <0.001 Рh <0.001 <0.05 <0.01 <0.05 <0.001 н.с. <0.05 н.с. <0.05 н.с. Table 9. The statistical significance of differences in allele frequencies (Pa) and observed heterozygosity (Ph) among Nipponacmea moskalevi settlements from polluted (Gaidamak Bay) and unpolluted (Srednyaya Bay) sites in Vostok Bay. Locus Ааt-1 Ааt-2 Gpi 1-Gphdh Idh-2 Lap Ме-1 Ме-2 Iph 6-Pgd Est-3 Mean Ра н.с. н.с. <0.001 н.с. н.с. н.с. н.с. н.с. н.с. <0.001 н.с. <<0.001 Рh н.с. н.с. <0.01 <0.05 <0.05 н.с. н.с. н.с. <0.01 н.с. н.с. н.с. I 0.999 0.997 0.950 0.999 0.999 0.999 0.998 1.000 1.000 0.983 0.998 0.993 Table 10. Percent of loci with statistically significant differences (P<0.05) in genotypic (G), allele (A) frequencies and observed heterozygosity (H) among conspecific settlements of marine mussels from polluted and unpolluted sites in the Peter the Great Bay (Sea of Japan) Species N G A H Nakhodka Bay (2) 15 13.3 13.3 6.6 Vostok Bay (2) 15 26.6 26.6 33.3 Ussuriyski Bay, Gould Horn Bay (2) 15 40.0 40.0 6.6 Amurski Bay (2) 15 20.0 20.0 3.3 Tegula rustica Vostok Bay, Amurski Bay (2) 8 50.0 50.0 25.0 Nipponacmea moskalevi Vostok Bay (2) 11 18.5 18.5 36.0 Ischnochiton hakodadensis Nakhodka Bay, Vostok Bay, Amurski Bay (3) 10 70.0 70.0 70.0 Mytilus trossulus, Locality CONCLUSIONS: We were found the statistically significant differences in allele frequencies and in level of observed heterozygosity at some loci by comparison settlements from polluted and unpolluted sites of 4 species marine mollusks in Peter the Great Bay. The greater part of observed differences were cause by rare genotypes and rare alleles. The per sent of losi which are subject to natural selection is quite high. An absents the differences in mean heterozygosity among mollusk settlements from polluted and unpolluted sites are not consistent with the hypothesis of selective advantage of high-heterozygosity genotypes under anthropogenic pollution stress. An estimations of differences in genotype and allele frequencies and in level of observed heterozygosity at some loci may be the variables for monitoring of quality sea environments. Our data are in agreement with the theory of the adaptive significance of allozyme polymorphism. THANK YOU FOR ATTENTION!